Periodic acceptor excitation spectroscopy of single molecules
- 6 February 2007
- journal article
- research article
- Published by Springer Science and Business Media LLC in European Biophysics Journal
- Vol. 36 (6), 669-674
- https://doi.org/10.1007/s00249-007-0133-7
Abstract
Alternating-laser excitation (ALEX) spectroscopy has recently been added to the single-molecule spectroscopy toolkit. ALEX monitors interaction and stoichiometry of biomolecules, reports on biomolecular structure by measuring accurate Förster resonance energy transfer (FRET) efficiencies, and allows sorting of subpopulations on the basis of stoichiometry and FRET. Here, we demonstrate that a simple combination of one continuous-wave donor-excitation laser and one directly modulated acceptor-excitation laser (Periodic Acceptor eXcitation) is sufficient to recapitulate the capabilities of ALEX while minimizing the cost and complexity associated with use of modulation techniques.Keywords
This publication has 18 references indexed in Scilit:
- Three-Color Alternating-Laser Excitation of Single Molecules: Monitoring Multiple Interactions and DistancesBiophysical Journal, 2007
- Shot-Noise Limited Single-Molecule FRET Histograms: Comparison between Theory and ExperimentsThe Journal of Physical Chemistry B, 2006
- A Single‐Molecule Förster Resonance Energy Transfer Analysis of Fluorescent DNA–Protein Conjugates for NanobiotechnologySmall, 2006
- Separating Structural Heterogeneities from Stochastic Variations in Fluorescence Resonance Energy Transfer Distributions via Photon Distribution AnalysisThe Journal of Physical Chemistry B, 2006
- Accurate single-pair Förster resonant energy transfer through combination of pulsed interleaved excitation, time correlated single-photon counting, and fluorescence correlation spectroscopyJournal of Biomedical Optics, 2006
- Probing structural heterogeneities and fluctuations of nucleic acids and denatured proteinsProceedings of the National Academy of Sciences of the United States of America, 2005
- Retention of Transcription Initiation Factor σ70 in Transcription Elongation: Single-Molecule AnalysisMolecular Cell, 2005
- Alternating-Laser Excitation of Single MoleculesAccounts of Chemical Research, 2005
- Comparison of Photophysical and Colloidal Properties of Biocompatible Semiconductor Nanocrystals Using Fluorescence Correlation SpectroscopyAnalytical Chemistry, 2005
- Fluorescence-aided molecule sorting: Analysis of structure and interactions by alternating-laser excitation of single moleculesProceedings of the National Academy of Sciences of the United States of America, 2004